Fig. 2: Cholesterol accumulation induces phagocytic fragility in monocyte-derived macrophages. | Nature Communications

Fig. 2: Cholesterol accumulation induces phagocytic fragility in monocyte-derived macrophages.

From: Oncolytic viruses engineered to enforce cholesterol efflux restore tumor-associated macrophage phagocytosis and anti-tumor immunity in glioblastoma

Fig. 2

a Quantification of intracellular cholesterol levels in cell subsets within intracranial GL261-bearing C57BL/6 J model. n = 9 mice per group. b Quantification of cholesterol levels in splenic macrophages and tumor macrophages within intracranial GL261-bearing C57BL/6J model and G422-bearing KM model. n = 5 mice per group. c Quantification of populations of centrally resident macrophages (microglia) and monocyte-derived TAMs during tumor progression. n = 5 mice per group. d Representative Giemsa staining and quantification of the cell size of peripheral splenic macrophages and TAMs. The upper images are representative of 2 spleen samples and 2 tumor samples. Scale bar, 20 μm. The lower FCM plots represent 5 spleen samples and 5 GL261 tumor samples. e Representative TEM images of human TAMs. The nuclei (N, upper left) of TAMs showed distinct heterochromatin clumps (dark); the primary (L1) and secondary (L2) lysosomes, and large, poorly cleared phagosomes (P) were observed within the cytoplasm. The images are representative of 2 samples. f Representative flow plots comparing phagocytosis-mediated cholesterol uptake by TAMs and BMDMs. n = 2 independent experiments. Quantification of ex vivo phagocytosis in TIF-treated (g) and cholesterol-treated (h) BMDMs. n = 5 biological samples. Experimental setup (i) and quantification of in vivo phagocytosis (j) in adoptively transferred cholesterol-treated macrophages. n = 7 mice per group. Quantification of cholesterol levels in CD11b+CD68+ human TAMs (k) and CD11b+F4/80+ mouse TAMs (l) with high or low phagocytosis-inhibitory receptors. Preparation of single-cell suspensions from patient GBM tumors (n = 6) and mouse GBM tumors (n = 7). m Representative immunofluorescence images of TAMs. F4/80 (green), Filipin III (blue), and Siglec-10 (red). Experiment conducted once, n = 3 samples. n Quantification of cholesterol levels and phagocytosis-inhibitory receptor expression in macrophages. Spleen, n = 6. Tumor, n = 7. Statistical significance was determined using the Mann–Whitney test (Two-tailed) in a, b, d, g, h, j, l, n, or the paired t test (Two-tailed) in k. Data shown are the mean ± SD. Source data are provided in the Source Data file.

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